Development of Salivary Gland Proteins

果蝇的激素反应胶基因在果蝇3龄后期的唾液腺中有高水平表达。与其他粘合基因不同的是，gs-4在发育早期和第二个组织——前脑室中表达。becendorf博士建议使用足迹、体外突变和种系转化来确定哪些顺式作用序列控制这种独特的表达模式。由于Sgs- 4增强子由多个部分可互换的元件组成，我们将研究这些元件的相互作用及其调控功能的差异。为了分离gs-4的反式调节因子，我们将在噬菌体lambda中构建针对唾液腺DNA结合蛋白的单克隆抗体文库。用增强子dna -蛋白复合物筛选后分离单克隆抗体。这些单克隆基因将用于筛选cDNA文库。分离的cDNA克隆和用于分离它们的抗体将用于表征新基因的调控和表达模式以及蛋白质的dna结合活性。

The hormone responsive glue genes of Drosophila are expressed at high levels in the salivary glands of late third instar larvae. Unlike the other glue genes, Sgs-4 is expressed at early times in development and in a second tissue, the proventriculus. Dr. Beckendorf proposes to use footprinting, in vitro mutatgenesis, and germ line transformation to establish which cis acting sequences govern this unique expression pattern. Because the Sgs- 4 enhancer is made up of multiple, partially interchangeable elements, the interactions of these elements and the differences in their regulatory functions will be examined. To isolate trans- regulators of Sgs-4, a monoclonal antibody library directed against salivary gland DNA binding proteins will be constructed in phage lambda. Monoclonal antibodies will be isolated after screening with enhancer DNa-protein complexes. These monoclonals will then be used to screen a cDNA library. Isolated cDNA clones and the antibodies used to isolate them will then be used to characterize the regulation and expression patterns of the new genes and the DNA-binding activities of the proteins.